title: Development of PCR-based method for detection of Enterobacteriaceae in septicemia.
abstract: OBJECTIVE
  Sepsis is a systemic inflammatory response associated with high mortality rates in the clinical setting. A multiplex endpoint polymerase chain reaction (PCR) based assay for rapid detection of enterobacteriaceae involved in septicemia, which included Internal Control (IC) and 16S rDNA, is presented here. To develop a panel of primers for DNA fragments of 16S rDNA, enterobacteriaceae, IC, and evaluate analytical sensitivity and specificity of the test.
  
  
  METHODS
  Primers for amplification of enterobacteriaceae, IC, and16S rDNA were designed, and then PCR was performed. Minimal analytical sensitivity was determined by cloning and colony PCR, and specificity was tested on the basis of their respective standard strains. This study is a cross-sectional Model.
  
  
  RESULTS
  Our results showed the rpoB gene as the most promising target for detection of enterobacteriaceae by PCR amplification. Specificity and sensitivity of endpoint PCR were 100%, 100%, and 100%, and 10, 1, and 100 copies/reaction for enterobacteriaceae, IC, and 16S rDNA, respectively.
  
  
  CONCLUSIONS
  The molecular panel presented offers the advantage of an easy, reliable, and cost-effective system when compared to other molecular detection methods. However, further evaluation is needed. Our assay holds promising for more rapid pathogens related in clinical sepsis.
authors.affiliation: Department of Microbiology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Fazzeli
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Arabestani
authors.forename: Bahram Nasr
authors.initials: BN
authors.lastname: Esfahani
authors.forename: Farzin
authors.initials: F
authors.lastname: Khorvash
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Pourshafie
authors.forename: Sharareh
authors.initials: S
authors.lastname: Moghim
authors.forename: Hajieh Ghasemian
authors.initials: HG
authors.lastname: Safaei
authors.forename: Jamshid
authors.initials: J
authors.lastname: Faghri
authors.forename: Tahmine
authors.initials: T
authors.lastname: Narimani
keywords: Blood culture; Intensive Care Unit; pathogenic bacteria; polymerase chain reaction; sepsis